To continue a sequel
on antibodies and characteristic difficulties, I would like to take a position concerning
independent validation. In 2012 the movement of independent validation commenced.
The initiative stands firm that the science we do now in hardly reproducible and
the products we use hopelessly rely on luck to work. Despite the trend to use
published products I believe validation of each product is necessary.

Researchers have
identified that around 75% of the research articles are not reproducible. Which
in turn means that 100% of products used in these articles are dubious. These
numbers show the lack of legitimacy in articles and products. Thankfully the
25% remain and the product used are accepted. They are accepted only because
the article has been successfully reproduced in a different laboratory, either
for a purpose of reproducibility or a follow-up research. Imagine if all
products would be independently validated. We would see a sudden increase in the
productivity of research. This will not only benefit your research but all the
research to come.

Furthermore, a validated product is solid in
an article for future references. Once you publish an article and indicate a
product that has been independently validated it attracts more attention, than
a research with non-validated products. It brings your results more legitimacy.
The future research to come will trust your article more, than the article of
someone else with a similar product.

Finally, by spending a little more on
independent validation you save funds on other resources needed to repeat the
experiment. Once you invest in independent validation it provides an insurance
for your product. Without this insurance a failed experiment due to a faulty
product prompts to consumption of valuable laboratory material. Unfortunately,
the only person that is to blame is you yourself. In Antibodychain, one of the
future projects includes option to independently validate every antibody from
every producer. This insurance will provide you with benefits of security and trustworthiness.

To conclude, today’s topic touches upon a currently
sensitive aspect of research. We need to validate our products since it allows
us to reduce failed experiments, comprise more validity for future referencing,
and save capital of investors, save laboratory equipment. It is better to be
safe than sorry, I recommend to have the validation mandatory.

In this blog entry I would like to focus on the classical
issue of retailers and manufacturers. Indeed, the field of life science cannot
leave the classical problem between two schools of thought: we need retailers
for customers or we need only manufacturers for products. To my opinion both
are important, but in today’s highly interconnected world the later takes the
lead.

Once the product has been in the
hands of a retailer he has the resources to market the product, pitch it to
customers, and create loyalty to the retailer. In return he creates a customer
basis for his producer on subjective basis. This works for many industries, but
for life sciences the paradigm is shifting. Internet of things allows us to
have the access to all information about life science products. The abundance
of information in the area allows us to be less subjective in terms of decision
making. This way we need less and less interaction with the sales men, less
marketing, less pitching, and less retail.

In addition, the attention is drawn
over advantages from the manufacturers themselves. They provide a longer
warranty, with direct answers from the producer. They do not add a margin of
handling costs to their final product. The producers are happy to customize the
form of the final product, they have the raw products.

Finally, chemicals or biological
agents are not sold over the counter. The limitation blinds the purchaser from
the final product he/she may receive if purchased from a salesman. Laboratories
over the Netherlands, Germany and Switzerland see this as an emerging trend.
The data sheet has to be present in front of the interested group, details
about safety, purity, usage is present as well. The internet and the
manufacturer have this covered. All the details are in their well-maintained
websites and are accessible to anyone.

All in all, chemicals, proteins,
antibodies, life science products are high importance group of materials that
shift to a different, more efficient, internet based presentation. They begin
to deviate from retailers since the access tomanufacturersthrough internet is cheaper and as
easy.

There is an ongoing discussion about antibody
validation, systematic antibody publication, and controlled, centralized
product identification. Academic discussion such as a webinar organized by
''The Scientist'' (An Urgent Need for Validating and Characterizing Antibodies)
has addressed these heated topics with emphases on antibodies alone. To my
opinion the trap that lies beneath us is that we are encouraged to trust a
product instead of a method.

First of all, methodologies regarding
antibody usage change with higher technological opportunities. Previously, a
cell sample could be incubated for a specific amount of time with the antibody,
you applied a washing buffer, added a secondary antibody, incubated it, washed
it again, dried it and took a look under a microscope. The simple idea has
advanced into washing methods involving TBST, Tween combinations, incubation
time differences and staining dye differences. Now every scientist can relate
that methods are not perfectly generalized, they are constantly improved and
modified. Progressively we are taught the contemporary way of blotting,
staining and counting cells. This means that the protocols handled by companies
can be and most likely will be different from the ones scientists have taught
each other in autonomous institutes. Scientists should start questioning which
company has optimized their staining methods. This way a research laboratory
can internalize the method of staining, blotting, counting to produce the same
result as indicated in the validated article.

Furthermore, publications indicate the
antibody used, but not the method to use. By looking at a research paper you
will likely end up with finding an antibody sold by a big company, which
associates itself with a product. While within a big company there are a lot of
small companies that actually produce and individually validate the antibody.
This means each company has a different method of testing which they use to
validate their antibody. Generally an antibody user lacks this information thus
user lacks methods used by a producer company. Once the user performs an
experiment with a product according to ‘big’ generic protocol, the antibody
performance can be questioned. We should use the exact same staining method of
a published article or the method developed by an antibody producer.

Unfortunately, it is easier to publish
an antibody than a method. It will take some time for scientific community to
recognise the importance of changing protocols. ''Generalized'' application
protocols circle around the websites of big biotech, pharma companies, but they
are clearly different, developed for their line of products. Users should be
more aware of what type of methods have been previously used by a company in
that publication. Some companies use vigorous testing methods, thus their
products are applicable in more scenarios, others do easy and cheap testing
thus you need to be careful.

All in all, published companies should
not be the flagman why we use antibodies of that particular article. The
article has to link company methods with your own methods. This will take more
time to find the right method, but the result will make the antibody experience
much better.

For decades there has
been a standard procedure to find antibodies of your own interest. A scientist
raises a hypothesis. The hypothesis postulates that certain proteins are
involved in a pathway. He retrieves literature to find the antibodies used, he
looks up on the internet for the company mentioned in the literature, and he
purchases the antibodies from the same company. Unfortunately the traditional
methods should have changed by now. Science reporters Vivien Marx, Jeffrey M.
Perkel, scientific advisors from Abcam and Origine agree. This one is going to
be a hard one to swallow: The antibodies production chains are not the same.
The same antibodies of the same company produced at different points of time
are different. The antibodies that you would order now are not the same
antibodies you have read about in the article. You put yourself in danger of
losing time.

The leading cause of
this is changing production circumstances. People within production chain
change, or engineers optimize the purification methods, or the animals for
immunization were raised differently. It is never perfect match. Every antibody
lot will have a slight difference whether these are polyclonal or monoclonal
antibodies. Each company struggles to keep their production line the same,
because their main short term goal is rarely the quality, but the quantity of
the product. For this simple reason the reproducibility of the antibody in
experiments will have deviation through time.

Every experiment
values time as the biggest opportunity cost and every failed run of Western
Blot or failed stain of Immunohistochemistry leads to great losses. For example
in the lab of Prof. Albert Heck, we could not more express our concern if the
antibody used for a Western Blot shows multiple bands instead of a single one
anticipated. You first point fingers at yourself, you test and retest believing
that the antibody you have purchased is according to the article, but in the
end of the day you lose more time instead of claiming an alternative antibody.
The great risk of unwanted differences between the moment the article was
published and the moment you are performing your experiment is widening
exponentially. Immunization of animals take place from a batch of independently
prepared antigens that go from hand to hand. This freeze thaw cycling
throughout the years influence the end antibody. Unfortunately each sample
antibody for immunization has small differences, which influence the response
of the tissue, there is close to no way to stop this.

In case people are
still confident in using the antibodies that have been published 6 months ago,
they are more than welcome to proceed, actually highly encouraged. The antibody
with the same lot number will surely bring you the same specificity, no off
target binding effect, will probably give you the same results. The only thing
you need to do is put some effort in finding this antibody.

All in all,
differences in antibody production is mirrored in the results of research
institutes. They plot the graphs according to the antibodies their producers
supplied, but not according to the past publications they have read. The antibody
industry still does not have the golden standard which could be followed in
order to save time and have 95% reproducible articles. Antibodychain is
tackling the motion and encourages others to compare their “ideal” products
with others at www.theantibodyshop.com.